During their metastatic spread, cancer cells need to remodel the extracellular matrix in order to migrate through stromal compartments adjacent to the primary tumor. Dissemination of breast carcinoma cells is mediated by membrane type 1-matrix metalloproteinase (MT1-MMP/MMP14), the main invadopodial matrix degradative component. Here, we identify MT1-MMP as a novel interacting partner of dual-specificity LIM Kinase-1 and -2 (LIMK1/2), and provide several evidence for phosphorylation of tyrosine Y573 in the cytoplasmic domain of MT1-MMP by LIMK. Phosphorylation of Y573 influences association of F-actin binding protein cortactin to MT1-MMP-positive endosomes and invadopodia formation and matrix degradation. Moreover, we show that LIMK1 regulates cortactin association to MT1-MMP-positive endosomes, while LIMK2 controls invadopodia-associated cortactin. In turn, LIMK1 and LIMK2 are required for MT1-MMP-dependent matrix degradation and cell invasion in a three-dimensional type I collagen environment. This novel link between LIMK1/2 and MT1-MMP may have important consequences for therapeutic control of breast cancer cell invasion.

SignificanceThis study shows that an oncogenic mammary epithelial cell surrounded by normal cells can extrude basally in vivo and invade surrounding tissues without formation of a primary tumor. Here, we show that overexpression of the key polarity protein atypical protein kinase C ι (aPKCi) is sufficient for triggering basally oriented epithelial cell extrusion and early cell invasion into the mammary gland stroma. Moreover, we highlight the importance of the difference between the mechanical properties of aPKCi-overexpressing cells and those of the normal surrounding cells associated with the decrease of vinculin at the cell junction, which triggers cell segregation, the first step toward promoting and controlling the direction of cell extrusion.